Dataset title: Trophic Interactions, Habitat Use, and Pollution Loads of Bottlenose Dolphins (Tursiops Truncatus) in the Florida Coastal Everglades, Florida, USA, 2013-2019 Dataset ID: doi:10.6073/pasta/4da8a26388dc34d5f86e26eeb74a4a84 Dataset Creator Name: Dr. Valeria Paz Position: Courtesy Post Doctoral Researcher Organization: Florida International University Email: vpaz003@fiu.edu Name: Dr. Michael Heithaus Position: Dean School of Arts and Sciences Organization: Florida International University Email: heithaus@fiu.edu Name: Dr. Jeremy Kiszka Position: Assistant Professor Organization: Florida International University Email: jkiszka@fiu.edu Name: Kirk Gastrich Position: Lab manager and researcher Email: kgastric@fiu.edu Dataset Abstract Cetaceans can feed at upper trophic levels and occur from freshwater to open-ocean ecosystems. Due to their abundance, mobility, and high metabolic rates, they have the potential to affect the structure and function of ecosystems through both top-down and bottom-up pathways. To better understand what ecological roles they may play in a system, it is important to understand patterns and drivers of their abundance, habitat use, and trophic interactions. I investigated the trophic interactions and pollutant exposure of common bottlenose dolphins (Tursiops truncatus) of the Florida Coastal Everglades. Based on bulk stable isotope analysis of tissue samples collected using biopsy sampling, it appears that despite their high mobility, bottlenose dolphins restrict their foraging within the habitats where they were sampled. Trophic position and foraging locations affected exposure to pollutants, with high levels of mercury found in dolphins estimated to forage at higher trophic levels and feeding within an inland bay. Mercury levels also varied with age and sex. Dolphins and their prey both contained substantial mercury levels and dolphins’ health could be impacted by this exposure, but the selenium levels we measured might counteract these negative effects. Geographic Coverage Bounding Coordinates Geographic description: Florida Coastal Everglades West bounding coordinate: -81.0707961 East bounding coordinate: -80.8018979 North bounding coordinate: 25.2600055 South bounding coordinate: 25.2071794 Temporal Coverage Start Date: 2013-02-27 End Date: 2019-02-22 Data Table Entity Name: 2023_LTER_DolphinBiopsy Entity Description: Bottlenose Dolphin Biopsy data- Isotopes, age, total mercury and selenium Object Name: 2023_LTER_Dolphin.csv Data Format Number of Header Lines: 1 Attribute Orientation: column Field Delimiter: , Number of Records: Attributes Attribute Name: Date Attribute Label: Date Attribute Definition: Date Storage Type: dateTime Measurement Scale: Missing Value Code: Attribute Name: Sample_ID Attribute Label: Sample_ID Attribute Definition: Sample ID name Storage Type: string Measurement Scale: text Missing Value Code: Attribute Name: Location Attribute Label: Location Attribute Definition: Location area of biopsy sample Storage Type: string Measurement Scale: Florida Bay= Florida Bay sample location Gulf of Mexico= Gulf of Mexico sampling location Shark River= Shark River sampling location Tarpon Bay= Tarpon Bay sampling location Whitewater Bay= Tarpon Bay sampling location Missing Value Code: Attribute Name: Season Attribute Label: Season Attribute Definition: season of time of the sampling event - wet or dry Storage Type: string Measurement Scale: Dry= Dry season- November to May Wet= Wet season- June to October Missing Value Code: Attribute Name: Blubber_Delta15N Attribute Label: Blubber_Delta15N Attribute Definition: Nitrogen Stable isotope of blubber tissue Storage Type: float Measurement Scale: Units: permil Number Type: real Missing Value Code: -9999 ( No sample available for analysis) Attribute Name: Blubber_Delta13C Attribute Label: Blubber_Delta13C Attribute Definition: Carbon Stable isotope of blubber tissue Storage Type: float Measurement Scale: Units: permil Number Type: real Missing Value Code: -9999 ( No sample available for analysis) Attribute Name: Skin_Delta15N Attribute Label: Skin_Delta15N Attribute Definition: Nitrogen Stable isotope of skin tissue Storage Type: float Measurement Scale: Units: permil Number Type: real Missing Value Code: Attribute Name: Skin_Delta13C Attribute Label: Skin_Delta13C Attribute Definition: Carbon Stable isotope of skin tissue Storage Type: float Measurement Scale: Units: permil Number Type: real Missing Value Code: Attribute Name: Latitude_DD Attribute Label: Latitude_DD Attribute Definition: Latitude Storage Type: float Measurement Scale: Units: Decimal Degrees Number Type: real Missing Value Code: Attribute Name: Longitude_DD Attribute Label: Longitude_DD Attribute Definition: Longitude Storage Type: float Measurement Scale: Units: Decimal Degrees Number Type: real Missing Value Code: Attribute Name: Sex Attribute Label: Sex Attribute Definition: Sex of individual sampled Storage Type: string Measurement Scale: F= Female M= Male Missing Value Code: Attribute Name: Age Attribute Label: Age Attribute Definition: Average estimated age Storage Type: float Measurement Scale: Units: Age unit Number Type: real Missing Value Code: -9999 ( No sample available for analysis) Attribute Name: low_age Attribute Label: low_age Attribute Definition: Low range of estimated range Storage Type: float Measurement Scale: Units: Age unit Number Type: real Missing Value Code: -9999 ( No sample available for analysis) Attribute Name: high_age Attribute Label: high_age Attribute Definition: High range of estimated age Storage Type: float Measurement Scale: Units: Age unit Number Type: real Missing Value Code: -9999 ( No sample available for analysis) Attribute Name: ppm_THg Attribute Label: ppm_THg Attribute Definition: Total Mercury ppm Storage Type: float Measurement Scale: Units: ppm Number Type: real Missing Value Code: -9999 ( No sample available for analysis) Attribute Name: THg_molar Attribute Label: THg_molar Attribute Definition: Total Mercury molar weight Storage Type: float Measurement Scale: Units: grams per mole Number Type: real Missing Value Code: -9999 ( No sample available for analysis) Attribute Name: Se_ppm Attribute Label: Se_ppm Attribute Definition: Selenium ppm Storage Type: float Measurement Scale: Units: ppm Number Type: real Missing Value Code: -9999 ( No sample available for analysis) Attribute Name: Se_molar Attribute Label: Se_molar Attribute Definition: Selenium molar weight Storage Type: float Measurement Scale: Units: grams per mole Number Type: real Missing Value Code: -9999 ( No sample available for analysis) Data Table Entity Name: 2023_LTER_Dolphin_Prey Entity Description: Prey targeted by bottlenose dolphins- Isotopes, total mercury, and selenium Object Name: 2023_LTER_Dolphin_Prey.csv Data Format Number of Header Lines: 1 Attribute Orientation: column Field Delimiter: , Number of Records: Attributes Attribute Name: Sample ID Attribute Label: Sample ID Attribute Definition: Sample ID name Storage Type: string Measurement Scale: text Missing Value Code: Attribute Name: Date Attribute Label: Date Attribute Definition: Date Storage Type: dateTime Measurement Scale: Missing Value Code: Attribute Name: Gear_Type Attribute Label: Gear_Type Attribute Definition: Gear type Storage Type: string Measurement Scale: CN= Cast Net HL= Hook and line- fishing pole Missing Value Code: Attribute Name: Location Attribute Label: Location Attribute Definition: Location of sampling Storage Type: string Measurement Scale: Florida Bay= Florida Bay sample location Shark River= Shark River sampling location Tarpon Bay= Tarpon Bay sampling location Whitewater Bay= Whitewater Bay sampling location Missing Value Code: Attribute Name: Latitude_DD Attribute Label: Latitude_DD Attribute Definition: Latitude Storage Type: float Measurement Scale: Units: Decimal Degrees Number Type: real Missing Value Code: Attribute Name: Longitude_DD Attribute Label: Longitude_DD Attribute Definition: Longitude Decimal Degrees Storage Type: float Measurement Scale: Units: Decimal Degrees Number Type: real Missing Value Code: Attribute Name: Species Attribute Label: Species Attribute Definition: Species Storage Type: string Measurement Scale: Caranx sp.= Caranx sp. C. nebulosus = Cynoscion nebulosus C. undecimalis= Centropomus undecimalis Caranx sp.= Caranx sp. E. saurus= Elops saurus L. griseus= Lutjanus griseus L. rhomboides= Lagodon rhomboides M. cephalus= Mugil cephalus S. ocellatus= Sciaenops ocellatus Missing Value Code: Attribute Name: Muscle_Delta15N Attribute Label: Muscle_Delta15N Attribute Definition: Nitrogen Stable isotope of muscle tissue Storage Type: float Measurement Scale: Units: permil Number Type: real Missing Value Code: Attribute Name: Muscle_Delta13C Attribute Label: Muscle_Delta13C Attribute Definition: Carbon Stable isotope of muscle tissue Storage Type: float Measurement Scale: Units: permil Number Type: real Missing Value Code: Attribute Name: Total_Length_cm Attribute Label: Total_Length_cm Attribute Definition: total length Storage Type: float Measurement Scale: Units: centimeter Number Type: real Missing Value Code: Attribute Name: Fork_Length_cm Attribute Label: Fork_Length_cm Attribute Definition: Fork Length Storage Type: float Measurement Scale: Units: centimeter Number Type: real Missing Value Code: -9999 (No Fork Length ) Attribute Name: Weight_kg Attribute Label: Weight_kg Attribute Definition: Weight Storage Type: float Measurement Scale: Units: kilogram Number Type: real Missing Value Code: -9999 (Not able to be measured) Attribute Name: THG_ppm Attribute Label: THG_ppm Attribute Definition: Total Mercury ppm Storage Type: float Measurement Scale: Units: ppm Number Type: real Missing Value Code: Methods Method Step Description Stable Isotope Analysis Dolphin skin samples were removed from the blubber layer using a methanol-rinsed stainless-steel blade by cutting as close to the interface between the two layers as possible without retaining any blubber. Dolphin samples (both skin and blubber) and prey samples (muscle tissue) were dried in an oven at 60 °C for 24-48 hr and then homogenized into a fine powder (DeNiro and Epstein, 1978). Lipids were removed only from dolphin samples using a 2:1 chloroform:methanol mixture for one minute with a solvent volume three times greater than the sample, left undisturbed for at least 45 min, centrifuged for 10 min, and then the supernatant was removed. After repeating this process three times, each dolphin sample was rinsed in deionized water and dried. The dried prey samples and lipid-extracted dolphin samples were then weighed, (0.4–0.7 mg) , and placed in a 4x6 mm tin capsule for SIA using a ThermoFinnigan Delta V isotope ratio mass spectrometer (IRMS) coupled with a NA 1500 NE elemental analyzer. Isotopic ratios are reported in the standard delta (δ) notation relative to the international standards of Vienna Pee Dee belemnite (δ13C) and atmospheric nitrogen (δ15N) using the following equation (1): (1) δ(‰) = [(RSample/RStandard) – 1] x 1000 All stable isotope analyses (SIA) were completed at the Southeast Environmental Research Center Stable Isotope Laboratory at Florida International University. Analytical reproducibility was based on replicates of internal standards including bovine liver and glycine; variation among standards was 0.12‰ and 0.10‰ for δ13C and δ15N, respectively. The mean skin and blubber C: N ratios were less than 3, indicating an adequate lipid extraction (Lesage et al., 2010). References: DeNiro, M. J., & Epstein, S. (1978). Influence of diet on the distribution of carbon isotopes in animals. Geochimica et cosmochimica acta, 42(5), 495-506. Lesage, A., Lelli, M., Gajan, D., Caporini, M. A., Vitzthum, V., Miéville, P., ... & Emsley, L. (2010). Surface enhanced NMR spectroscopy by dynamic nuclear polarization. Journal of the American Chemical Society, 132(44), 15459-15461. Method Step Description DNA extraction and sex identification In order to sex sampled individual dolphins, DNA was extracted from skin tissue (< 2 mm) using a modified phenol/chloroform protocol from Sambrook et al. (1989) and described in detail in Beal et al. (2019). Concentrations of extracted DNA were checked by nanodrop and quality checked by gel electrophoresis. Aliquots at 30 ng/µL concentration were created for sex and age analysis procedures. Sex was identified by PCR and gel electrophoresis using the following PCR protocol: 10 µL H2O, 0.2 µL Thermo ScientificTM DreamTaqTM (5 U/ µL), 4 µL DreamTaq Polymerase Buffer, 1 µL dNTPs (2mM), 1 µL MgCl2 (25mM), 1 µL of Forward and Reverse ZFX primers (1 µM) and Forward SRY primer, and 0.5 µL Reverse SRY primer (1 µM) to which 1.5 µL of aliquoted DNA was added for a total volume of 21.2 µL for each sample. Thermocycler settings were as follows: initial denaturation of 95°C for 30s, followed by 30 cycles of 94°C for 30s, 51°C for 45s, and 72°C for 45s, then a final extension at 68°C for 5 min, ending with a final hold at 4°C. PCR products were run on a 2.5% agarose gel with Smart Glow pre-stain at 90 Voltz for approximately 1.5 hours and viewed with UV light. References: Beal, A. P., Kiszka, J. J., Wells, R. S., & Eirin-Lopez, J. M. (2019). The bottlenose dolphin epigenetic aging tool (BEAT): A molecular age estimation tool for small cetaceans. Frontiers in Marine Science, 6, 561. Sambrook, J., Fritsch, E. F., & Maniatis, T. (1989). Molecular cloning: a laboratory manual (No. Ed. 2). Cold spring harbor laboratory press. Method Step Description Age was estimated for all individuals using the Bottlenose Epigenetic Aging Tool (BEAT; Beal et al. 2019). Extracted DNA was divided into 600 ng aliquots (20ng/µL) and subjected to bisulfite transformation and pyrosequencing using the established protocol for the BEAT (EpigenDx, Hopkinton, MA, USA). The % DNA methylation was used in the BEAT model equation (2) to estimate age for all individuals. The BEAT model was determined to have a root mean square error of 5.14 when cross validated using the leave one out cross validation (LOOCV) method. This means that individual age estimates produced by the BEAT model are within this error range (Beal et al., 2019). Using this potential error, both high and low age estimates were also used in analyses. High age estimates were used for this study. (2) Age estimate= 8.602 + 4.392 (%DNA GRIA_5) – 1.149 (%DNA TET2_2) References: Beal, A. P., Kiszka, J. J., Wells, R. S., & Eirin-Lopez, J. M. (2019). The bottlenose dolphin epigenetic aging tool (BEAT): A molecular age estimation tool for small cetaceans. Frontiers in Marine Science, 6, 561. Distribution Online distribution: https://pasta.lternet.edu/package/data/eml/knb-lter-fce/1243/1/467e5f71379c6e948adee9b37e482a2f Intellectual Rights These data are embargoed so that the graduate student who collected them will have a chance to publish their findings before the data can be accessed by others. Dataset Keywords biopsies mercury total mercury carbon isotopes nitrogen isotopes aging DNA methylation bottlenose dolphins fish stable isotopes Tursiops truncatus isotopes stable isotopes fishes Dataset Contact Name: Kirk Gastrich Position: Lab manager and researcher Organization: Florida International University Email: kgastric@fiu.edu Name: Dr. Valeria Paz Position: Courtesy Post Doctoral Researcher Organization: Florida International University Email: vpaz003@fiu.edu Name: Michael Heithaus Position: Dean School of Arts and Sciences Organization: Florida International University Email: heithaus@fiu.edu